I only have 16 ul of 263.6 ng/ul RNA. I currently have 2.72 units/ul of DNAse I, but wondering how much DNAse to use and if I add DNAse to my 16 ul RNA, how long should I incubate at 37C? Additionally, what is the best way to inactivate the DNAse without degrading my RNA? Any suggestions would be greatly appreciated.
look up the prortocol for the DNase I you have, it should have calculations for how many units of DNase I to use by concentration of sample. Recognize that, depending on how you measured your RNA concentration, some portion of it at least is DNA, so expect to get a smaller amount back than you start with.
In my opinion; DNA will be inactivated by heating but this can damage your RNA. So, you can use EDTA to inactivate DNaseI and try LiCl precipitation to remove DNAse.
Hi
the best way is not to try eliminating DNA but to design primers at the right positions, I mean almost one primer designed on an exons junction, so that only RNA will be taken as matrix.
fred
Give your sample a DNASE Treatment. Dnase comes in a kit maretted by various companies like thermo fisher.
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